title: HER-2 gene amplification in human breast cancer without concurrent HER-2 over-expression.
abstract: BACKGROUND
  Testing for human epidermal growth factor receptor-2 (HER-2) in breast cancer is performed by either immunohistochemistry (IHC) or in situ hybridization (ISH). The growth factor receptor-bound protein-7 (GRB7) gene is in close proximity to HER-2 on chromosome 17q11-12 and codes a signal transduction molecule shown to be an independent adverse marker in breast cancer.
  
  
  METHODS
  HER-2 and GRB7 protein expression from 613 frozen breast tumors was determined by Western analysis. HER-2 protein results were confirmed with IHC. Commercial HER-2 FISH was performed on a subset of tumors with multi-probe FISH used to assess the extent of HER-2 gene amplification. mRNA expression was determined by Multi-plex RT-PCR.
  
  
  RESULTS
  Seven tumors with GRB7 protein over-expression scored HER-2 FISH amplified but had no HER-2 protein over-expression. Four of the 7 tumors showed elevated GRB7 but not HER-2 mRNA over-expression. The breast cancer cell line HCC3153 did not over-express HER-2 protein but showed HER-2 FISH amplification of a limited segment around the HER-2 gene. Ten breast cancer tumors from the TCGA database had gene copy number increases around HER-2 without HER-2 mRNA or protein over-expression.
  
  
  CONCLUSIONS
  A subset of human breast cancers that test positive with FISH for HER-2 gene amplification do not over-express HER-2 protein. One mechanism for this discordance is the incomplete amplification of the smallest HER-2 region of chromosome 17q11-12, which includes GRB7. HER-2 gene amplification without protein over-expression is clinically significant because patients with such tumors are unlikely to benefit from HER-2 targeted therapy.
authors.affiliation: Division of Hematology and Medical Oncology, Oregon Health and Science University, 3181 SW Sam Jackson Park Road, MC L586, Portland, OR 97239 USA ; Portland VA Medical Center, 3710 SW US Veterans Hospital Road, Portland, OR 97239 USA ; Knight Cancer Institute, Oregon Health and Science University, 3181 SW Sam Jackson Park Road, Portland, OR 97239 USA.
authors.forename: Shiuh-Wen
authors.initials: SW
authors.lastname: Luoh
authors.forename: Betsy
authors.initials: B
authors.lastname: Ramsey
authors.forename: Amy
authors.initials: A
authors.lastname: Hanlon Newell
authors.forename: Megan
authors.initials: M
authors.lastname: Troxell
authors.forename: Zhi
authors.initials: Z
authors.lastname: Hu
authors.forename: Koei
authors.initials: K
authors.lastname: Chin
authors.forename: Paul
authors.initials: P
authors.lastname: Spellman
authors.forename: Susan
authors.initials: S
authors.lastname: Olson
authors.forename: Edward
authors.initials: E
authors.lastname: Keenan